Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
GRASP65 Double Nickase Plasmid (h): sc-402436-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GRASP65 Double Nickase Plasmid (h) consists of a pair of plasmids each encoding a D10A mutated Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed to knockout gene expression with greater specificity than its CRISPR/Cas9 KO counterpart
- Paired gRNA sequences are offset by approximately 20 bp to allow for specific Cas9-mediated double nicking of the genomic DNA, which mimics a DSB
- One plasmid in the pair contains a puromycin-resistance gene for selection; the other plasmid in the pair contains a GFP marker to visually confirm transfection
- GRASP65 Double Nickase Plasmid (h) and GRASP65 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GORASP1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: GRASP65 Antibody (D-12): sc-374423
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GRASP65 Double Nickase Plasmid (h) | sc-402436-NIC | 20 µg | $410.00 | |||
GRASP65 Double Nickase Plasmid (h2) | sc-402436-NIC-2 | 20 µg | $410.00 |
GORASP1 encodes GRASP65, a peripheral membrane Golgi protein that helps maintain Golgi stack architecture and supports cisternal stacking and ribbon formation through regulated oligomerization and interactions with golgins and vesicle-tethering factors. GRASP65 contributes to Golgi reassembly after mitosis and participates in membrane trafficking and protein sorting along the secretory pathway, influencing glycosylation-dependent processing of cargo. Its phosphorylation-dependent dynamics couple Golgi organization to cell-cycle progression and stress responses that remodel secretory compartments. Altered Golgi structure and trafficking programs involving GRASP65 have been associated with pathological states characterized by disrupted proteostasis and aberrant secretion, including neurodegeneration and cancer-related cell biology.
GRASP65 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GORASP1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GORASP1. When directed to adjacent sites on opposite DNA strands, the two nickases generate offset single-strand nicks that together produce a staggered double-strand break, requiring coordinated on-target activity from both guides. The resulting DNA break is resolved by endogenous cellular repair pathways, most commonly through non-homologous end joining (NHEJ), leading to insertions or deletions that disrupt GORASP1 function. By requiring dual sgRNA engagement at the target locus, the double nicking approach enhances editing specificity and provides a complementary CRISPR strategy for applications where additional control over targeting precision is desired.
To support efficient identification of edited cells, one plasmid encodes GFP for fluorescent visualization of transfected populations, while the companion plasmid carries a puromycin resistance gene for antibiotic selection. Together, these features support efficient enrichment of co-transfected populations and simplify the validation of GORASP1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.